Novel Broad-Spectrum Aptamer Used to Build Fe–Co Nanoporous Carbon Highly Sensitive Sensing Platform for the Detection of Multiple Organophosphorus Pesticides

In this work, broad-spectrum aptamers for organophosphorus pesticides (OPs) were obtained by alternate target systematic evolution of ligands by exponential enrichment screening. The secondary and tertiary structure analyses of the aptamer inferred that the neck-loop structure formed a G-triplex structure with the target. In addition, optimization of the sheared aptamer resulted in a stronger affinity (Kd = 86.74 nM), which was increased by 2 orders of magnitude compared to similar aptamers. A novel electrochemical biosensor was prepared by modifying an aptamer labeled with an electroactive substance (methylene blue) on the surface of nanoporous carbon containing Fe–Co (Fe–Co/NPC). When a target bound to the aptamer, a G-triplex structure was formed close to the electrode surface. The aptamer phosphate backbone labeled with methylene blue enhanced the electron-transfer efficiency and resulted in signal changes. The biosensor exhibited an excellent sensitivity (7.32 fM) and a wide detection range (1 × 10–13 to 1 × 10–3 M) for OPs under optimal conditions, enabling simultaneous detection of multiple OPs in vegetables.